Plant derived seed extract rich in essentially fatty acids derived from Salvia hispanica L. seed: composition of matter, manufacturing process and use

ABSTRACT

A  Salvia hispanica  L. derived seed oil extract composition of matter containing from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of ALA to LA, 4-10% of C-18 mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8% of C-16 saturated fatty acid in a mixed triglyceride form stable at room temperature of 12-24 months containing a mixture of selected antioxidants.

RELATED APPLICATION(S)

This application is a continuation application of Ser. No. 13/206,757filed Aug. 10, 2011, which is a divisional application of Ser. No.12/419,321 filed Apr. 7, 2009, which is based upon prior filedprovisional application Ser. No. 61/043,773 filed Apr. 10, 2008, thedisclosures of which are incorporated by reference in their entirety.

FIELD OF THE INVENTION

This invention relates to plant-derived seed extracts and methods ofmaking same.

BACKGROUND OF THE INVENTION

It is well known in the literature that Polyunsaturated Fatty Acids(PUFAs) of all types are highly susceptible to peroxide, free radicaland light induced degradation including rancification and polymerizationmaking them unsuitable for human consumption. For example, it is wellknown that flax seed oil, also known as linseed oil, readily undergoesfree radical oxidation to advantageously form polymeric surfacesincluding oil based paints, hard furniture finishes and linoleumflooring. In addition, many companies offer flax seed oil for humanconsumption as a dietary supplement or food ingredient because of thehigh levels of PUFAs found in raw flax seed and its expeller pressedoils and more particularly Alpha-Linolenic Acid (ALA) and Linolenic Acid(LA). Many flax seed oil product labels suggest that the product must berefrigerated at all times due to the instability of such PUFAs in flaxseed oil. Careful examination of the majority of commercially availableflax seed oils obtained by expeller pressing, including those typicallystored under refrigerated conditions, unfortunately reveals that theyare unfit for human use based on their measured Peroxide Values (PVs).Such PV values above 3 meq/Kg (milliequivalents/gram) are deemed notsuitable for salad oil applications and PV values above 10 meq/Kg aredeemed to be unsuitable for human use because the measured PV value maybe an early indicator of rancidity and free radical induced degradation.On the other hand, PV values taken alone do not adequately characterizesuch oils since a low PV value can also be associated with PUFA's thathave already gone through the rancification process. Typical testing hasrevealed flax seed oil products sold for human consumption with observedPV's as high as 130 meq/kg also characterized with the odor associatedwith short chain aldehydes that make such oils “rancid” to olefactorysenses.

In fact, most raw seed based oils in common cooking and baking use, suchas soybean, corn and canola seed oils naturally contain enough PUFAsmaking them unsuitable, without further processing, for use as cookingoils. Therefore unless such PUFA containing raw seed oils arehydrogenated to fully saturated triglycerides using hydrogen and acatalyst prior to their use in cooking applications, they are consideredto be unfit for use as cooking oils. These oils are typically firstisolated by, for example, expeller pressing the appropriate seed,filtration of the crude seed oil to remove biomass and the resultingoil, containing significant levels of PUFAs, is then catalyticallyhydrogenated to reduce the PUFA content to levels suitable for use ofthe resulting oil in cooking applications. If the hydrogenation processis incomplete, however, the resulting mixtures are found to contain bothundesirable heat labile PUFAs that quickly undergo rancification tosmall chain aldehydes in the resulting heated cooking oil as well asunsaturated trans-fatty acids which are believed to be detrimental toanimal and especially human health.

Therefore, those skilled in the art will recognize the great difficultyin producing a shelf stable PUFA mixture wherein the PUFA content is ashigh as 70% wt/wt of the resulting seed extract from a natural seedsource that then exhibits extraordinary room temperature stability.

SUMMARY OF THE INVENTION

In accordance with a non-limiting aspect, a composition of matterincludes a supercritical CO₂ Salvia hispanica L. derived seed oilcomprising from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of C-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and comprising a mixture ofselected antioxidants.

In another aspect, the composition of matter includes dietary supplementingredients such as docosahexaenoic acid (DHA) and/or eicosapentaenoicacid (EPA) in pectin or gelatin based confectionary dietary supplementdelivery systems. EPA, DHA, docosahexaenoic acid (DPA) orgamma-linlolenic acid (GLA), fish oil, krill oil, krill oil concentrate,borage oil, evening primrose oil, olive oil or other plant, animal oralgal based seed or fruit oils are admixed therein either alone or incombination. Lipophilic antioxidants are added either alone or incombination with a) phenolic antioxidants including at least one ofsage, oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d)carotenoids including at least one of astaxanthin, lutein, andzeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g) Butylatedhydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i) TertiaryButyl hydroquinone (TBHQ).

In yet another aspect a hydrophilic antioxidant or sequesterant includeshydrophilic phenolic antioxidants including at least one of grape seedextract, tea extracts, ascorbic acid, citric acid, tartaric acid, andmalic acid.

A method of manufacturing and method of using the composition is alsoset forth.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention will now be described more fully hereinafter withreference to the accompanying drawings, in which preferred embodimentsof the invention are shown. This invention may, however, be embodied inmany different forms and should not be construed as limited to theembodiments set forth herein. Rather, these embodiments are provided sothat this disclosure will be thorough and complete, and will fullyconvey the scope of the invention to those skilled in the art.

In accordance with a non-limiting aspect, a room temperature, shelfstable mixture of an approximate 3.3:1 mixture of alpha-linolenic acid(“ALA”, “Omega-3 polyunsaturated fatty acid” (“PUFA”)) to linoleic acid(“LA”, “Omega-6 PUFA) has been prepared in the presence of limitedamounts of saturated and mono-unsaturated fatty acids as their mixedtriglycerides by the use of either supercritical fluid CO₂ solventextraction of pre-milled Salvia hispanica L. seed alone, and moreparticularly, supercritical fluid CO₂ solvent extraction in the presenceof mixtures of hydrophilic and lipophilic antioxidants, or, by the useof a common organic solvent extraction such as hexane or by the use ofexpeller pressing techniques.

Such Omega-3 and Omega-6 PUFAs are well known as essential fatty acidsin man and many animals, which are useful in humans and animals inpromoting, for example, a heart healthy condition in man. It is alsowell known, however, that PUFAs are extremely susceptible to rapid,uncontrollable free radical mediated degradation.

A PUFA rich seed oil extract is prepared from Salvia hispanica L. seedwhich contains one of nature's most favorable seed based concentrationsand ratios of the essential fatty acids and more specifically theessential fatty acids ALA and LA in a ratio of approximately 3.3:1 andmore particularly a mixture of said ALA and LA that is stable at roomtemperature for long periods of time when appropriately treated withantioxidants either before, during or after, or any combination thereof,supercritical fluid extraction, hexane extraction or expeller pressingof the seeds.

Such oils are used either alone or advantageously in combination withother ingredients, for example, algae, plant or fish derivedalpha-linolenic acid (ALA) or linoleic acid (LA) metabolites such aseicosapentaenoic acid (EPA), docosapentaenoic acid (DPA),gamma-linlolenic acid (GLA) or docosahexaenoic acid (DHA) or anycombination thereof, incorporated into appropriate foods, beverages ordietary supplements for the prevention or mitigation of such diseases ascardiovascular disease, arthritis, pain, blood clotting, dry eyes andbrain health.

Such disease mitigation has been associated with the competitive controlof the LA metabolic cascade and the resulting metabolic cascade productsfrom LA metabolism known commonly as eicosanoids, such as the series 2and 3 prostaglandins and thromboxanes, the series 4 leucotrienes andlipoxins and the series 5 leuotrienes all of which are potent plateletaggregators and/or inhibitors, pro-inflammatories, vasodilators,bronchoconstrictors, or anti-asthmatics and the like.

The consumption of ALA has been shown to be a very effective competitivesubstrate of delta-6 desaturase, which is known to be the rate limitingenzymatic step in both ALA and LA metabolism to the metabolic productsdiscussed above.

Attempted extraction of Saliva hispanica L. un-milled seed, usingsupercritical CO₂ even at extraordinarily high pressures of 1000 bar orhexane solvent at atmospheric pressures, yields very little, if any,seed oil, therefore the seed must be milled prior to extraction. Theextent of the milling, as measured by particle size distribution, isadvantageous to the extraction process in accordance with a non-limitingaspect since the higher the surface area, the higher will be theefficiency and completeness of the extraction process by either organicsolvent based or supercritical fluid based processes. In addition, it isadvantageous to mill the seed in a blanket of inert gas such as nitrogento prevent per-oxidative processes from taking place that wouldotherwise be initiated in the presence of air or oxygen and light.

In one embodiment, Salvia Hispanica L. whole seed is either firstcommutated in a standard knife or hammer mill or more preferably rollermilled, preferably under a cold nitrogen atmosphere, to produce crackedseed biomass. The biomass is preferably treated with one or morehydrophilic and/or lipophilic antioxidants by mixing the antioxidants tothe resulting biomass. In another embodiment, the antioxidant may beadvantageously added to the seed prior to or during the milling processor at the point of extraction without pre-blending said antioxidantsevenly throughout the resulting biomass due to the nature of theextraction process. The biomass is then transferred to a supercriticalfluid extraction unit for separation of the seed oil from the cracked orflake-rolled biomass.

Alternatively, the pre-prepared biomass can be transferred to a commonhexane solvent extractor, or an expeller press for example, and the oilextracted from the biomass accordingly. Preferably either process isconducted in the absence of oxygen or air.

The supercritical fluid extraction of the milled seed admixed withhydrophilic and/or lipophilic antioxidants is accomplished by subjectingthe pre-milled cracked or flake-rolled seed to supercritical CO₂, or CO₂and propane as a co-solvent or supercritical propane alone at from40-1000 bar at from 30-100 Deg. C. More preferably the seed oil isextracted from the biomass between 50-800 bar at 50-90 deg. C. in suchCO₂ amounts measured in kgs/kg of biomass and for such times as may berequired to extract large portions of the seed oil content from thebiomass. In addition, entrainment solvents can be added to thesupercritical fluid to further enhance the efficiently of suchextractions. For example, supercritical carbon dioxide extraction of thebiomass can be enhanced by the addition of propane to the supercriticalextraction fluid.

The resulting seed oil dissolved in supercritical solvent(s) is nextallowed to fractionate in two separate pressure step-down stagesallowing the collection of a light and heavy fraction of seed oilextract. This light fraction also contains water that has beenco-extracted from the seed mass. The resulting seed oil, afterdegassing, is separated from any water that may have been carried overduring the extraction of the biomass containing said water. The lightfraction of the seed oil extract is rich in taste and odor componentsand may be admixed with the heavy fraction or may be discarded dependingon the desired product characteristics.

After separation of the water remaining in each fraction, the fractionsare then held under nitrogen or other inert gas and additional amountsof lipophilic and/or hydrophilic antioxidants may then be added. Inaddition, the resulting fractions may also be treated with bleachingclay, carbon and such other processing aids as may be required to renderthe oil suitable for its intended use in humans and animals.

The PV of the resulting seed oil extract is typically under 2.0 meq/Km,while accelerated decomposition, using a Rancimat instrument, remarkablyindicates an extrapolated room temperature shelf life of from 1-2 years.When the same process is repeated without the use of antioxidants, theresulting PV is surprisingly under 10.0 meq/Kg most probably due to theuse of supercritical CO2 resulting in minimal exposure of the oil tooxygen species, however the resulting oil quickly begins to buildperoxide value in the presence of air even when stored at temperaturesof 0 Degs. C. In addition, such unstabilized oils, under acceleratedrancimat testing exhibit very poor stability to heat and oxygen unlikethe rancimat performance observed in stablizied oils derived from theprocess described above.

The resulting supercritical fluid seed oil extract of the inventioncontains from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of ALA:LA,4-10% of C18 mono-unsaturated fatty acid, 1-5% of C-18 saturated fattyacid and 4-8% C-16 saturated fatty acid composition in a mixedtriglyceride form depending on the seed source employed.

On the other hand, if the process described above is conducted withoutthe use of hydrophilic and/or lipophilic antioxidants, the resultingseed oil extract exhibits an initial low PV but accelerated stabilitytesting using a Rancimat instrument indicates an extrapolated roomtemperature shelf stability of less than two months.

The stability of the resulting oil at room temperature that ismanufactured without the use of added antioxidants cannot be easilyexplained because of the available levels of the powerful naturalantioxidants found in Salvia hispanica L. whole seed whose activity canbe easily measured in Oxygen Radical Absorbance Capacity (ORAC) units.Salvia hispanica L. has a measured ORAC number of 3000 micromoles TEORAC units/gram of seed and is known to contain such antioxidants asmyricetin, quercetin, kaempferol, caffeic acid, and chlorogenic acid. Inaddition, it is well known that the Salvia hispanica L. whole seed,unlike many other seeds bearing PUFA containing oil, exhibits a shelflife of at least 5 years due to its structure and the naturallyoccurring antioxidants available in the seed matrix.

In addition, cold pressing of Salvia hispanica L. whole seed alsoproduces unstable seed oil without careful addition of appropriateantioxidants to the seed prior to the expeller pressing process.

In a non-limiting example the composition of matter is formed from asupercritical CO₂ derived Salvia hispanica L. derived seed oilcomprising from 60-88% PUPAS in a ratio of from 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of C-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and comprising a mixture ofselected antioxidants.

It includes docosahexaenoic acid (DHA) and/or eicosapentaenoic acid(EPA) in pectin or gelatin based confectionary dietary supplementdelivery systems and in another aspect EPA, DHA, docosapentaenoic acid(DPA) or gamma-linlolenic acid (GLA), fish oil, krill oil, krill oilconcentrate, borage oil, evening primrose oil, olive oil or other plant,animal or algal based seed or fruit oils are admixed therein.

Lpophilic antioxidants are added either alone or in combination with a)phenolic antioxidants including at least one of sage, oregano, androsemary; b) tocopherol, c) tocotrienol(s), d) carotenoids including atleast one of astaxanthin, lutein, and zeaxanthin; e) ascorbylacetate; f)ascorbylpalmitate g) Butylated hydroxytoluene (BHT); h) DocosapentaenoicAcid (BHA) and i) Tertiary Butyl hydroquinone (TBHQ). The hydrophilicantioxidant or sequesterant includes hydrophilic phenolic antioxidantsincluding at least one of grape seed extract, tea extracts, ascorbicacid, citric acid, tartaric acid, and malic acid in another aspect.

A method of manufacturing a Salvia hispanica L. derived seed oil inanother non-limiting example is set forth. The seed oil comprises from60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of alpha-linolenic acid(ALA) to linoleic acid (LA), 4-10% of C-18 mono-unsaturated fatty acid,1-5% of C-18 saturated fatty acid and 4-8% of C-16 saturated fatty acidin a mixed triglyceride form that is stable at room temperature for12-24 months and includes antioxidants. The method includes milling orroller press flaking Salvia hispanica L. seed in the absence of oxygento obtain a desired particle size distribution with or without theaddition of hydrophilic or lipophilic antioxidants during the particlesizing process. The resulting biomass is subjected to supercriticalfluid CO₂ extraction in the presence of lipophilic and or hydrophilicantioxidants. Any resulting seed oil fractions are collected. The wateris separated in each fraction.

Any resulting seed oil fractions can be treated with additionalantioxidants to afford a desired room temperature stability. The extentof oil extraction can be controlled by particle size distribution of themilled or flaked seed. Propane can be added in mixture withsupercritical CO₂ in the supercritical state as an extraction solvent.In yet another aspect solvent can be extracted using hexane extractionat or near atmospheric pressures and the resulting boiling point ofhexane in the absence of oxygen, separating the resulting water from theoil/hexane mixture and removing the hexane solvent by distillation at orbelow atmospheric pressure in the absence of oxygen.

Lipophilic antioxidants can be added to increase the room temperaturestability of the resulting oil. The lipophilic antioxidants can be addedeither alone or in combination with a) phenolic antioxidants includingat least one of sage, oregano, and rosemary; b) tocopherol, c)tocotrienol(s), d) carotenoids including at least one of astaxanthin,lutein, and zeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g)Butylated hydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA) and i)Tertiary Butyl hydroquinone (TBHQ). The resulting dewatered seed oil canbe treated with bleaching clay or activated carbon.

Pre-milled or roller press flaked seed can be treated with a lipophilicor hydrophilic antioxidant(s) prior to solvent extraction. Thehydrophilic antioxidant or sequesterant can be formed from hydrophilicphenolic antioxidants including at least one of grape seed extract, teaextracts, ascorbic acid, citric acid, tartaric acid, and malic acid.

A method of mitigating or preventing cardiovascular disease, arthriticpain and inflammation, platelet aggregation, or treating dry eyesyndrome, pre-menstrual symptoms or modifying immune response in humansor animals is set forth by applying an effective amount of a dietarysupplement, food or beverage to which has been a composition mixedtherewith and comprising a Salvia hispanica L. derived seed oilcomprising from 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 ofalpha-linolenic acid (ALA) to linoleic acid (LA), 4-10% of C-18mono-unsaturated fatty acid, 1-5% of C-18 saturated fatty acid and 4-8%of C-16 saturated fatty acid in a mixed triglyceride form that is stableat room temperature for 12-24 months and includes antioxidants.

In one aspect, an emulsifying agent is added. In another aspect, nano-and/or micro-particles of rice or sugarcane based polycosanol aredispersed for enhancing a heart healthy dietary supplement. A stabilizedoil in a fruit juice concentrate, fruit puree or other water basedflavoring is dispersed in the presence of maltodextrin, or othercarbohydrates, and a suitable emulsifying or emulsion stabilizationagent that is vacuum spray dried to form an amorphous or crystallinesolid suitable for use as a flavoring ingredient carrying stabilizedPUFAs in flavored dietary supplements, foods and beverages. In yetanother aspect, oil based flavors and fragrances suitable for use as aningredient in foods, beverages and cosmetics are added. ALA and LA arealso added as essential fatty acids.

Many modifications and other embodiments of the invention will come tothe mind of one skilled in the art having the benefit of the teachingspresented in the foregoing descriptions and the associated drawings.Therefore, it is to be understood that the invention is not to belimited to the specific embodiments disclosed, and that themodifications and embodiments are intended to be included within thescope of the dependent claims.

That which is claimed is:
 1. A composition of matter comprising asupercritical CO₂ fluid Salvia hispanica L. derived seed oil comprisingfrom 60-88% PUFAs in a ratio of from 3.1:1-3.3:1 of alpha-linolenic acid(ALA) to linoleic acid (LA), 4-10% of C-18 mono-unsaturated fatty acid,1-5% of C-18 saturated fatty acid and 4-8% of C-16 saturated fatty acidin a mixed triglyceride form that is shelf stable at room temperaturefor 12-24 months and comprising an effective amount of added lipophilicand/or hydrophilic antioxidant stabilizer as a preservative that extendsthe shelf life of the seed oil, wherein the Salvia hispanica L. derivedseed oil is made by subjecting the Salvia hispanica L seed tosupercritical fluid CO2 extraction to produce a seed oil extract;fractionating the resulting seed oil extract in separate pressurestep-down stages for collecting light and heavy fractions of seed oilextract; and separating the heavy fraction from the light fraction toform the final seed oil from the heavy fraction.
 2. The composition ofmatter according to claim 1, and further comprising docosahexaenoic acid(DHA) and/or eicosapentaenoic acid (EPA) in pectin or gelatin basedconfectionary dietary supplement delivery systems.
 3. The composition ofmatter according to claim 1, and further comprising EPA, DHA,docosapentaenoic acid (DPA) or gamma-linlolenic acid (GLA), fish oil,krill oil, krill oil concentrate, borage oil, evening primrose oil,olive oil or other plant, animal or algal based seed or fruit oilsadmixed therein.
 4. The composition of matter according to claim 1, andfurther comprising lipophilic antioxidants either alone or incombination with a) phenolic antioxidants including at least one ofsage, oregano, and rosemary; b) tocopherol, c) tocotrienol(s), d)carotenoids including at least one of astaxanthin, lutein, andzeaxanthin; e) ascorbylacetate; f) ascorbylpalmitate g) Butylatedhydroxytoluene (BHT); h) Docosapentaenoic Acid (BHA), and i) TertiaryButyl hydroquinone (TBHQ).
 5. The composition of matter according toclaim 1, and further comprising a hydrophilic antioxidant orsequesterant comprising hydrophilic phenolic antioxidants including atleast one of grape seed extract, tea extracts, ascorbic acid, citricacid, tartaric acid, and malic acid.
 6. The composition of matteraccording to claim 1, wherein a peroxide value of the seed oil is under2.0 meq/Km.
 7. A composition of matter comprising a supercritical CO₂fluid Salvia hispanica L. derived seed oil comprising from 60-88% PUFAsin a ratio of from 3.1:1-3.3:1 of alpha-linolenic acid (ALA) to linoleicacid (LA), 4-10% of C-18 mono-unsaturated acid, 1-5% of C-18 saturatedfatty acid and 4-8% of C-16 saturated fatty acid in a mixed triglycerideform that is shelf stable at room temperature for 12-24 months andcomprising an effective amount of added lipophilic and/or hydrophilicantioxidant stabilizer as a preservative that extends the shelf life ofthe seed oil, wherein the Salvia hispanica L. derived seed oil is madeby subjecting the Salvia hispanica L seed to supercritical fluid CO2extraction to produce a seed oil extract; fractionating the resultingseed oil extract in separate pressure step-down stages for collectinglight and heavy fractions of seed oil extract; and separating the heavyfraction from the light fraction to form the final seed oil from theheavy fraction.
 8. The composition of matter according to claim 7, andfurther comprising docosahexaenoic acid (DHA) and/or eicosapentaenoicacid (EPA) in pectin or gelatin based confectionary dietary supplementdelivery systems.
 9. The composition of matter according to claim 7, andfurther comprising EPA, DHA, docosapentaenoic acid (DPA) orgamma-linlolenic acid (GLA), fish oil, krill oil, krill oil concentrate,borage oil, evening primrose oil, olive oil or other plant, animal oralgal based seed or fruit oils admixed therein.
 10. The composition ofmatter according to claim 7, and further comprising lipophilicantioxidants either alone or in combination with a) phenolicantioxidants including at least one of sage, oregano, and rosemary; b)tocopherol; c) tocotrienol(s); d) carotenoids including at least one ofastaxanthin, lutein, and zeaxanthin; e) ascorbylacetate; f)ascorbylpalmitate; g) Butylated hydroxytolene (BHT); h) DocosapentaenoicAcid (BHA); or i) Tertiary Butyl hydroquinone (TBHQ).
 11. Thecomposition of matter according to claim 7, and further comprising ahydrophilic antioxidant or sequesterant comprising hydrophilic phenolicantioxidants including one of grape seed extract, tea extracts, ascorbicacid, citric acid, tartaric acid, and malic acid.
 12. The composition ofmatter according to claim 7, wherein a peroxide value of the seed oil isunder 2.0 meq/Km.